Isolation and Analysis of Small RNAs from Virus-Infected Plants

In this chapter, we detail some of the methods available to the researcher for isolating and analyzing virus-derived small RNAs (vsRNAs). These methods have been successfully used for four plant viruses: Cucumber mosaic virus (CMV), including the CMV Y-Satellite, Turnip mosaic virus (TuMV), Potato leaf roll virus (PLRV), and Tomato spotted wilt virus (TSWV) from inoculated Arabidopsis thaliana plants (Fusaro et al. EMBO Rep 7:1168–1175, 2006; Curtin et al. FEBS Lett 582:2753–2760, 2008). The protocols presented here can also be employed for the isolation of non-virus related small RNAs such as microRNAs (miRNAs) and hairpin RNA (hpRNA).